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Erythrocyte Lysis Reagent and Its Application

A red blood cell lysing and erythrocyte technology, applied in the direction of biological testing, microbial dissolution, instruments, etc., can solve the problems of insufficient lysis effect, insufficient lysis efficiency, poor purity of separated cells, etc., and achieve the ideal effect of lysis effect

Active Publication Date: 2021-02-23
GUANGDONG WESAIL BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, many conventional erythrocyte lysing reagents in the prior art have defects that the lysing effect is not good enough in practical applications, for example, the purity of the isolated cells is poor, or the lysing efficiency is not high enough, etc.

Method used

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  • Erythrocyte Lysis Reagent and Its Application
  • Erythrocyte Lysis Reagent and Its Application
  • Erythrocyte Lysis Reagent and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] 1 preparation method

[0077] Prepared according to the following concentration

[0078] -NH 4 Cl 0.15M,

[0079] -NaCl0.03M,

[0080] - sucrose 5w / v%,

[0081] -EDTA 0.1nM,

[0082] -Triton X-100 1v / v%,

[0083] -SDS 8w / v%, and

[0084] -NaHCO 3 50mM.

[0085] The preparation method is as follows: add each component into ultrapure water, dilute to 1L, mix well and dissolve, adjust pH to 8.0 with HCl or NaOH.

[0086] 2 applications

[0087] Whole blood samples were selected and tested by fluorescent immunochromatography at different lysis times, and the reference system was BioRad D-10 high performance liquid chromatography.

[0088] Take 10 μL of whole blood, add it to 490 μL of lysate, invert and mix for 5-300 s, and the lysate can obtain a clear and transparent liquid, then perform chromatography on the glycosylated hemoglobin immunochromatographic test strip, and read the test with a fluorescence detector result.

[0089] 3 Effect Evaluation

[0090] ...

Embodiment 2

[0093] 1 preparation method

[0094] Prepared according to the following concentration

[0095] -NH 4 Cl 0.4M,

[0096] -NaCl 0.01M,

[0097] - Trehalose 1w / v%,

[0098] -EDTA·Na 2 0.5nM,

[0099] -Tween 20 10v / v%,

[0100] -SDS 8w / v%, and

[0101] -NaHCO 3 5mM.

[0102] The preparation method is as follows: add each component into ultrapure water, dilute to 1L, mix well and dissolve, adjust pH to 8.5 with HCl or NaOH.

[0103] 2 applications

[0104] Since the detection of glycosylated hemoglobin is based on the complete lysis of red blood cells, the comparison of clinical data between the detection of glycosylated hemoglobin chromatography and high performance liquid chromatography can indirectly reflect the completeness of lysis and the stability of the effect of the lysate.

[0105] Select 64 fresh whole blood samples of glycosylated hemoglobin calibrated by BioRad D10, take 10 μL of whole blood each, add to 490 μL lysate, invert and mix for 10 seconds, and th...

Embodiment 3

[0107] 1 preparation method

[0108] Prepared according to the following concentration

[0109] -NH 4 Cl 0.25M,

[0110] - NaCl 0.02M,

[0111] - sucrose 3w / v%,

[0112] -EDTA·K 2 0.3nM,

[0113] -Tween 20 5v / v%,

[0114] -SDS 4w / v%, and

[0115] -NaHCO 3 30mM.

[0116] The preparation method is as follows: add each component into ultrapure water, dilute to 1L, mix well and dissolve, adjust pH to 8.2 with HCl or NaOH.

[0117] The lysate A and the lysate of the present invention were used to compare the lysate of glycated hemoglobin.

[0118]Among them, the formula and configuration method of lysate A are the same as the above-mentioned components in Example 3, the only difference is that the ingredients do not contain sucrose and SDS.

[0119] 2 applications

[0120] Lysis time

[0121] A) 50-fold lysing, lysing by shaking upside down at the same strength for 10 seconds, observing that there are still flaky red blood cells remaining in the lysate A, and the lys...

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Abstract

The invention relates to the field of biotechnology, in particular to a erythrocyte lysing reagent and its application. The erythrocyte lysis reagent is an aqueous solution containing the following components: NH 4 + / NH 3 、Na + , sucrose and / or trehalose, chelating agent, Triton X‑100 and / or Tween 20, SDS. The lysis time of the reagent is short, the lysis is complete, and the clinical compliance rate is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a erythrocyte lysing reagent and its application. Background technique [0002] Red blood cells, also known as red blood cells (RBC), are the most numerous blood cells in the blood, and are also the most important medium for transporting oxygen through the blood in vertebrates, and also have immune functions. Red blood cells contain hemoglobin, which makes blood red. However, when modern clinical diagnostic methods are used for diagnostic testing of blood samples, red blood cells interfere with many diagnostic tests, which requires the separation of red blood cells from blood samples before further testing of the samples. [0003] In the detection of blood samples, the presence of red blood cells will interfere with many diagnoses, and they need to be separated from the blood samples first. Generally, the method of first lysing and then separating is used; when the content of red bl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/50G01N33/53G01N33/532G01N33/558G01N33/569G01N33/80C12N1/06
CPCC12N1/06G01N33/5005G01N33/5306G01N33/532G01N33/558G01N33/56966G01N33/80
Inventor 刘丽萍于鸫迟晨
Owner GUANGDONG WESAIL BIOTECH CO LTD